Journal: The FASEB Journal

Article Title: Endothelial Exostosin‐1 Is Needed for Glycocalyx Integrity and Inflammatory Resolution in Acute Lung Injury

doi: 10.1096/fj.202601250R

Figure Lengend Snippet: Prolonged inflammatory cell accumulation in Ext1‐deficient mice during lipopolysaccharide (LPS)‐induced lung injury. (A) Immunohistochemical staining for CD11b + cells at 0–48 h after LPS administration. (B) Quantification of CD11b + cells. Ext1‐deficient mice (red line) show higher CD11b + cell retention at 48 h, indicating delayed inflammatory resolution. (C) Immunohistochemical staining for F4/80 + macrophages. (D) Quantification of F4/80 + cells. Cre + Ext1 flox/flox mice show persistent and increased macrophage accumulation, consistent with impaired inflammatory clearance. Data represent mean ± standard error of the mean (SEM). * p < 0.05 vs. Cre − controls.

Article Snippet: Sections were incubated with primary antibodies against CD11b (Abcam, ab16667; 1:200), F4/80 (Bioss, bs‐10701R; 1:200), or heparan sulfate (10E4 clone; Amsbio, #370255; 1:100) overnight at 4°C.

Techniques: Immunohistochemical staining, Staining

Journal: Journal of Clinical and Translational Hepatology

Article Title: Single-cell RNA Sequencing Analysis Reveals That Targeting PLG–PLGRKT Signaling-mediated Pro-fibrotic Scar-associated Macrophages Ameliorates Liver Fibrosis in Biliary Atresia

doi: 10.14218/JCTH.2026.00278

Figure Lengend Snippet: (A) Schematic illustration of Trem2 antibody–siRNA conjugates used for Plgrkt knockdown in SAMs. (B) Uptake of Cy5-labeled Trem2–siRNA conjugates by RAW264.7 macrophages observed by fluorescence microscopy. (C) In vivo localization of Cy5-labeled Trem2–siRNA conjugates with F4/80-Cy3-positive macrophages in liver tissue. (D) Schematic overview of the BDL mouse model and treatment groups. (E) Western blot analysis of Plg-RKT protein levels in liver tissues from different experimental groups. (F) H&E and Sirius Red staining of liver sections from each group. (G) Representative flow cytometry analysis of SAMs (Cd45 + F4/80 + Trem2 + Cd9 + ) in liver tissue. (H) Quantification of SAM frequencies across experimental groups. (I) mRNA expression levels of SAM-associated genes ( Trem2 and Cd9 ) measured by RT-qPCR. (J) mRNA expression levels of fibrosis-related genes ( Spp1 , Anxa2 , Gpnmb , Lgals1 , and Lgals3 ) measured by RT-qPCR. Data represent n = 5 per group. * P < 0.05, ** P < 0.01, and *** P < 0.001. BDL, bile duct ligation; H&E, hematoxylin and eosin; RT-qPCR, reverse transcription quantitative polymerase chain reaction; SAMs, scar-associated macrophages; siRNA, small interfering RNA; PLGRKT, plasminogen receptor with a C-terminal lysine.

Article Snippet: Non-parenchymal cells (1 × 10 6 cells) were Fc-blocked with TruStain FcX (BioLegend, San Diego, CA, USA) and stained with CD45 PE/Cy7 (BioLegend, 103113), F4/80 PerCP/Cy5.5 (Elabscience, E-AB-F0995UJ; Houston, TX, USA), TREM2 PE (BioLegend, 824805), CD9 APC (BioLegend, 124811), and Ly6G BV421 (BioLegend, 127627)( ).

Techniques: Knockdown, Labeling, Fluorescence, Microscopy, In Vivo, Tissue, Western Blot, Analysis, Staining, Flow Cytometry, Expressing, Quantitative RT-PCR, Ligation, Reverse Transcription, Real-time Polymerase Chain Reaction, Small Interfering RNA

Journal: Frontiers in Immunology

Article Title: UPF1-dependent glycolysis driven by LINC00887 facilitates macrophage M2 polarization in clear cell renal cell carcinoma

doi: 10.3389/fimmu.2026.1833383

Figure Lengend Snippet: LINC00887/UPF1-mediated aerobic glycolysis leads to lactic acid accumulation and facilitates immune evasion in ccRCC in vivo , A syngeneic tumor model was developed by injecting BALB/c mice with Renca cells expressing sh-NC or sh-LINC00887, followed by treatment with sh-NC or sh-UPF1 lentivirus. A-C: Assessment of tumor size (A) , volume (B) , and weight (C) . (D, E) qRT-PCR analyses for LINC00887 (D) and UPF1 (E) mRNA levels. (F) IHC analysis for UPF1, LDH-A, KI67 protein levels. (G) Kit assay for lactate levels in tumors. (H) Proportion of M2 macrophages (F4/80+CD206+) by flow cytometry. (I) Immunofluorescence assay for CD206+ macrophages. (J) Proportion of CD8 + T cells (CD3 + CD8 + ) in tumor tissues. ns, no significant difference. * p < 0.05. cRCC, Clear cell renal cell carcinoma; UPF1, Up-frameshift protein 1; LDH-A, Lactate dehydrogenase A; CD206, Mannose receptor C-type 1; CD3, Cluster of differentiation 3; CD8, Cluster of differentiation 8.

Article Snippet: Antibodies included: FITC Mouse monoclonal to CD68 (ab134351, abcam, UK), Pacific Blue anti-human CD86 Antibody (305417, yajimall, China), CD206 (MMR) Monoclonal Antibody (19.2)-PE (12-2069-42, ThermoFisher, USA), PerCP/Cyanine5.5 Anti-Mouse F4/80 Antibody (E-AB-F0995J, Elabscience, China), PE Anti-CD206 mouse antibody (E-AB-F1135D, Elabscience, China), APC Anti-mouse CD3 antibody (E-AB-F1013E, Elabscience, China), PE Anti-mouse CD8 alpha antibody (E-AB-F1104D, Elabscience, China).

Techniques: In Vivo, Expressing, Quantitative RT-PCR, Analysis, Flow Cytometry, Immunofluorescence